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- Table of Contents
NEFL · Western blot design guide
NEFL Western Blot Planning Guide
Plan a NEFL Western blot around the catalog-observed 61.5 kDa band, image-backed A02482-1 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
NEFL Western Blot Experimental Design Guide
Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | 61.5 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Hippocampus | |
| Negative control | Adipose tissue |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 61.5 kDa |
| ⓘ | Localization | Cell projection, axon / Cytoplasm, cytoskeleton |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated NEFL Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Cerebral cortex |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, standard transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A02482-1 at datasheet starting dilution |
| Primary incubation | Overnight at 4 °C with gentle agitation |
| Secondary | Species-matched HRP conjugate at validated dilution |
| Wash | 3 × 5 min in TBST |
| Detection | Chemiluminescent substrate |
| Exposure | Bracket exposures to avoid saturation |
Section 2
What Is the Expected NEFL Western Blot Band Size?
Use the product-observed 61.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 61.5 kDa | Matches the authoritative product WB observation. |
| 61.5 kDa calculated | Use as UniProt context, not as a replacement observed band. |
| Unexpected additional signal | Do not assign identity without orthogonal positive/negative controls. |
💡Expected NEFL appearancePlan around 61.5 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 61.5 kDa; use this as the primary experimental expectation. |
| Calculated mass | 61.5 kDa from UniProt P07196; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A02482-1. |
| Situation | Likely cause | Next action |
|---|---|---|
| 61.5 kDa | Matches the authoritative product WB observation. | Confirm with orthogonal controls and the linked product record. |
| Additional band | May reflect processing, modification, or non-specific signal. | Run a dilution series and compare positive/negative controls. |
| Weak signal | Target abundance or transfer may be limiting. | Verify transfer, increase positive-control abundance, and bracket exposure. |
Experiment evidence
Western blot images showing NEFL band
A02482-1 · NEFL
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NEFL Western blot
🧪Use Hippocampus as the first positive-control candidate and Adipose tissue as the HPA Not detected negative candidate.
Positive control: Hippocampus (High)
Negative control: Adipose tissue (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for NEFL
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Hippocampus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebral cortex | Reported tissue cells | High | Protein (HPA) | HPA → |
| Cerebellum | Reported tissue cells | Medium | Protein (HPA) | HPA → |
| Soft tissue | Reported tissue cells | Low | Protein (HPA) | HPA → |
Section 3
Advanced NEFL Western Blot Tips
Deeper troubleshooting and optimisation questions for NEFL, answered from its protein features.
Which band should guide the blot?
Use 61.5 kDa, the observation attached to the authoritative A02482-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 61.5 kDa expectation.
Which positive control should I start with?
Start with Hippocampus, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Adipose tissue as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for NEFL Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02482-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC12078432, PMC6249855, PMC9636496.
Boster reagents
NEFL Western Blot Reagents
Human/Mouse/Rat-reactive NEFL Western blot reagents with authoritative product imagery.
Real WB data
Anti-NEFL/NF-L Antibody Picoband®
Cat # A02482-1
Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.
Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.
References
- UniProt P07196
- Human Protein Atlas — NEFL
- A02482-1 product record
- PMC12078432 — NEFL Modulates NRN1-Mediated Mitochondrial Pathway to Promote Diacetylmorphine-Induced Neuronal Apoptosis.
- PMC6249855 — Integration of transcriptome and proteome profiles in glioblastoma: looking for the missing link.
- PMC9636496 — NEFL is overexpressed and it modulates invasion and migration in neuroendocrine-like PC3-ML2 prostate cancer cells.